Breeding method for transgenic mouse with conditional knockout of lncRNA DLX6-os1

Abstract

The invention relates to a breeding method for a transgenic mouse with conditional knockout of lncRNA DLX6-os1. The purpose is effectively achieved that the transgenic mouse as an animal model adaptsto different tissue organs for studying the effects of the lncRNA. A Dlx6-os1 conditional knockout carrier, the Dlx6-os1 gene, a transcript Dlx6-os1-201 ENSMUST00000159568.5 and three exons located ata chromosome 6 of the mouse are constructed, and the three exons are used as a conditional knockout area; a BAC clone of RP24-276P7 and RP24-260F14 in a C57BL / 6 mouse gene library is used as a template for PCR operation; in the purpose carrier, a self-deleting anchoring locus is inserted beside an Neo locus, an loxP locus is inserted beside the gene knockout area, and DTA negative selection is conducted; a gene mediated with a Cre enzyme is recombined, and a gene knockout plasmid carrier is obtained; the gene knockout plasmid carrier is transferred into an ES cell and hybridized with multipletransgenic mice with the Cre enzyme in different organs, and positive mice with the reduced expression level of the DLX6-os1 in different organs are obtained. The method is suitable for studying theeffects of the lnc RNA in different tissue organs, and a new way for diabetes diagnosis and treatment is developed.

Description

technical field [0001] The present invention relates to medical biology, in particular to a breeding method for conditional knockout lncRNA DLX6-os1 transgenic mice. Background technique [0002] In recent years, studies have shown that long non-coding RNA (Long non-coding RNA, LncRNA) is closely related to diabetic nephropathy, but there are few related research reports, and there is no report on the expression profile of LncRNA in patients with diabetic nephropathy, which cannot clarify the involvement of LncRNA in diabetic nephropathy. The pathogenesis of diabetic nephropathy, let alone its important position in the occurrence and development of diabetic nephropathy. [0003] Utilizing the characteristic biological sample bank of kidney disease, the high-throughput chip Arraystar human LncRNA chip V3.0 was used to screen the blood, urine and kidney tissue samples of diabetic nephropathy patients (confirmed by renal biopsy), and compare them with the normal control group. ...

AI Technical Summary

Problems solved by technology

At present, the commonly used method in the world is to inject the virus carrying the lncRNA into the corresponding tissues and organs in an attempt to change the expression level of the lncRNA, but there are many problems: 1. The infection efficiency of the virus is low, and it is impossible to effectively change the expression level of the lncRNA in specific organs The effect of expression level

2. Some lncRNAs have long sequences, which are not suitable for packaging viruses or cause low titers of viruses

3. Some organs are not suitable for virus infection and injection

[0005] Therefore, the C57 mice were genetically modified by genetic engineering technology to breed FLOX-DLX6-os1 transgenic mice. This mouse can be crossed with a variety of transgenic mice with cre enzymes in different organs to obtain reduced levels in different organs. DLX6-os1 expression level mice, these mice can be applied to different disease models, providing a very good animal model for clarifying the role of lncRNA DLX6-os1 in different tissues and organs, effectively solving the problem and applicable to different tissues and organs Research on the function of this lncRNA, but there is no public report so far

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